Description of the core: The Immunology core is located in the Kresge Eye Institute (KEl) building (Room 452) in dedicated space (Appendix: Floor plans of core space) and will provide immunological and flow cytometry expertise with priority of use to NEI R01 funded vision scientists. The new core which is fully accessible to all its users, will provide the following services. Laboratory services: ? Isolation of peripheral blood mononuclear cells (PBMC) for staining, cell sorting, or viable cryopreservation. ? Isolation of mucosal mononuclear cells (MMC) from mucosal fissure biopsies. ? Evaluation of cytokines and chemokines from aqueous and vitreous humor, corneal and retinal lysates and tissue culture supernatants by ELISA. ? ELISPOT assays for cell associated cytokine production. ? Morphological and biochemical assays for antigen-induced cell death. ? Flow Cytometry services (Accuri 6C and software): ? Immunophenotyping of whole blood, PBMC, and aqueous and vitreous fluids for: NK, DCs, Macrophages, B and T cell percentages, and their subsets, including maturation, trafficking, senescence, activation, in vivo proliferation and chemokine receptor expression. ? Flow cytometric functional assays, including NK cell function, antigen-specific CD4+ and CD8+ T cell cytokine expression, degranulation and proliferation. Evaluation of cell signaling by Phosflow using monoclonal antibodies specific for phosphorylated proteins. ? Flow Cytometer Based Cytokine Bead Array. ? Analysis of Intracellular cytokine expression in permeabilized cells. ? DNA/Cell cycle analysis. ? Apoptosis assays.